De novo characterization of the banana root transcriptome and analysis of gene expression under Fusarium oxysporum f. sp. Cubense tropical race 4 infection.
Bananas and plantains (Musa spp.) are among the most important crops in the world due to their nutritional and export value. However, banana production has been devastated by fungal infestations caused by Fusarium oxysporum f. sp. cubense (Foc), which cannot be effectively prevented or controlled. Since there is very little known about the molecular mechanism of Foc infections; therefore, we aimed to investigate the transcriptional changes induced by Foc in banana roots. We generated a cDNA library from total RNA isolated from banana roots infected with Foc Tropical Race 4 (Foc TR 4) at days 0, 2, 4, and 6. We generated over 26 million high-quality reads from the cDNA library using deep sequencing and assembled 25,158 distinct gene sequences by de novo assembly and gap-filling. The average distinct gene sequence length was 1,439 base pairs. A total of 21,622 (85.94%) unique sequences were annotated and 11,611 were assigned to specific metabolic pathways using the Kyoto Encyclopedia of Genes and Genomes database. We used digital gene expression (DGE) profiling to investigate the transcriptional changes in the banana root upon Foc TR4 infection. The expression of genes in the Phenylalanine metabolism, phenylpropanoid biosynthesis and alpha-linolenic acid metabolism pathways was affected by Foc TR4 infection. The combination of RNA-Seq and DGE analysis provides a powerful method for analyzing the banana root transcriptome and investigating the transcriptional changes during the response of banana genes to Foc TR4 infection. The assembled banana transcriptome provides an important resource for future investigations about the banana crop as well as the diseases that plague this valuable staple food.


Hardware-Based Hopfield Neuromorphic Computing for Fall Detection.
With the popularity of smart wearable systems, sensor signal processing poses more challenges to machine learning in embedded scenarios. For example, traditional machine-learning methods for data classification, especially in real time, are computationally intensive. The deployment of Artificial Intelligence algorithms on embedded hardware for fast data classification and accurate fall detection poses a huge challenge in achieving power-efficient embedded systems. Therefore, by exploiting the associative memory feature of Hopfield Neural Network, a hardware module has been designed to simulate the Neural Network algorithm which uses sensor data integration and data classification for recognizing the fall. By adopting the Hebbian learning method for training neural networks, weights of human activity features are obtained and implemented/embedded into the hardware design. Here, the neural network weight of fall activity is achieved through data preprocessing, and then the weight is mapped to the amplification factor setting in the hardware. The designs are checked with validation scenarios, and the experiment is completed with a Hopfield neural network in the analog module. Through simulations, the classification accuracy of the fall data reached 88.9% which compares well with some other results achieved by the software-based machine-learning algorithms, which verify the feasibility of our hardware design. The designed system performs the complex signal calculations of the hardware's feedback signal, replacing the software-based method. A straightforward circuit design is used to meet the weight setting from the Hopfield neural network, which is maximizing the reusability and flexibility of the circuit design.


The perception of South African adolescents regarding primary health care services.
Most of the South African public health facilities fail to provide adolescent-friendly health services. A quantitative, descriptive research study was conducted at Stinkwater, a rural area in Hammanskraal, South Africa. The objective of the study was to describe the adolescent's preferences regarding primary health care services. A survey was conducted among 119 adolescents. It was found that adolescents wished to be involved in the planning of the activities of the adolescent health service, and that friendliness and respect for adolescents were seen as desirable characteristics of an adolescent-friendly health care service. Adolescents preferred services to be available throughout the week and to be located at the school, youth center, community center, hospital, or clinic. Health education was indicated as a priority and the health care team should include different members of a multidisciplinary team. Adolescents preferred that their health services be separated from adult services and that a male nurse be employed in the adolescent service in order to create a less feminine image. It was also recommended that all adolescents be educated about the types of services available. Understanding health care service preferences of adolescents is needed in order to deliver optimal health care to this group.


Multiplexing PKA and ERK1&2 kinases FRET biosensors in living cells using single excitation wavelength dual colour FLIM.
Monitoring of different signalling enzymes in a single assay using multiplex biosensing provides a multidimensional workspace to elucidate biological processes, signalling pathway crosstalk, and determine precise sequence of events at the single living cell level. In this study, we interrogate the complexity in cAMP/PKA-MAPK/ERK1&2 crosstalk by using multi-parameter biosensing experiments to correlate biochemical activities simultaneously in time and space. Using a single excitation wavelength dual colour FLIM method we are able to detect fluorescence lifetime images of two donors to simultaneously measure PKA and ERK1&2 kinase activities in the same cellular localization by using FRET biosensors. To this end, we excite two FRET donors mTFP1 and LSSmOrange with a 440 nm wavelength and we alleviate spectral bleed-through associated limitations with the very dim-fluorescent acceptor ShadowG for mTFP1 and the red-shifted mKate2 for LSSmOrange. The simultaneous recording of PKA and ERK1&2 kinase activities reveals concomitant EGF-mediated activations of both kinases in HeLa cells. Under these conditions the subsequent Forskolin-induced cAMP release reverses the transient increase of EGF-mediated ERK1&2 kinase activity while reinforcing PKA activation. Here we propose a validated methodology for multiparametric kinase biosensing in living cells using FRET-FLIM.


Bond Strength and Flexural Capacity of Normal Concrete Beams Strengthened with No-Slump High-Strength, High-Ductility Concrete.
This study investigates the flexural behavior of normal-strength concrete (NSC) beams that were strengthened with no-slump, high-strength, high-ductility concrete (NSHSDC). A set of slant shear tests was performed to investigate the initial performance of the NSC substrate strengthened with NSHSDC. Slant shear tests considered two types of roughness of interface and five angles of the interface between NSC and NSHSDC. The test results showed that except for specimens with a 75° interface angle, the specimens with high roughness were conformed to the properties (14-21 MPa for 28 days) of the ACI Committee 546 recommendation. For flexural strength tests, NSC beams strengthened with an NSHSDC jacket on the top and bottom sides, three sides, and four sides resulted in strength increments of about 8%, 29%, and 40%, respectively, compared to the beams without NSHSDC jacket. Therefore, the use of NSHSDC is an effective method to improve the performance of NSC beams and is recommended for strengthening reinforced concrete members.


Sex differences in the effects of acute stress on behavior in the ultimatum game.
Acute stress affects human decision making. It has been argued that there are systematic sex differences in behavioral responses to acute stress, with males showing a 'fight or flight' and females showing a 'tend and befriend' response. A 'tend and befriend' response would suggest that women become more cooperative under acute stress, while men do not. We investigated the effects of acute stress on social behavior. We induced stress via the Trier Social Stress Test (TSST) and then immediately after measured how participants reacted to offers made in the ultimatum game by a male proposer. We found that female participants were less likely to reject offers under stress (n = 25) vs. no stress (n = 37), p = 0.009, independent of how fair these offers were, cooperative behavior consistent with the 'tend and befriend' hypothesis. Male participants when stressed (n = 30) did not show differences in rejections rates compared to the control condition (n = 26), p = 0.41. Our results provide support for a qualitatively different behavioral response to acute stress among men and women.


MYC activation cooperates with Vhl and Ink4a/Arf loss to induce clear cell renal cell carcinoma.
Renal carcinoma is a common and aggressive malignancy whose histopathogenesis is incompletely understood and that is largely resistant to cytotoxic chemotherapy. We present two mouse models of kidney cancer that recapitulate the genomic alterations found in human papillary (pRCC) and clear cell RCC (ccRCC), the most common RCC subtypes. MYC activation results in highly penetrant pRCC tumours (MYC), while MYC activation, when combined with Vhl and Cdkn2a (Ink4a/Arf) deletion (VIM), produce kidney tumours that approximate human ccRCC. RNAseq of the mouse tumours demonstrate that MYC tumours resemble Type 2 pRCC, which are known to harbour MYC activation. Furthermore, VIM tumours more closely simulate human ccRCC. Based on their high penetrance, short latency, and histologic fidelity, these models of papillary and clear cell RCC should be significant contributions to the field of kidney cancer research.


Ultrastructural demonstration of spirochetal antigens in synovial fluid and synovial membrane in chronic Lyme disease: possible factors contributing to persistence of organisms.
To perform the first systematic electronmicroscopic (EM) and immunoelectron microscopy (IEM) study of the pathological changes and the evidence of spirochete presence in synovial membranes and synovial fluid (SF) cells of patients with chronic Lyme arthritis. EM examination was performed on four synovial membrane and eight SF cell samples from eight patients with chronic Lyme disease. Spirochetal antigens in the samples were sought by IEM using monoclonal antibody to Borrelia burgdorferi outer surface protein A (OspA) as the immunoprobe. Prominent ultrastructural findings were surface fibrin-like material, thickened synovial lining cell layer and signs of vascular injury. Borrelia-like structures were identified in all four synovial membranes and in two of eight SF cell samples. The presence of spirochetal antigens was confirmed by IEM in all four samples studied (one synovial membrane and three SF cell samples). OspA labelling was in perivascular areas, deep synovial stroma among collagen bundles, and in vacuoles of fibroblasts in synovial membranes; and in cytophagosomes of mononuclear cells in SF cell samples. Electron microscopy adds further evidence for persistence of spirochetal antigens in the joint in chronic Lyme disease. Locations of spirochetes or spirochetal antigens both intracellulary and extracellulary in deep synovial connective tissue as reported here suggest sites at which spirochaetes may elude host immune response and antibiotic treatment.


Evaluation of the sensitivity of a pLDH-based and an aldolase-based rapid diagnostic test for diagnosis of uncomplicated and severe malaria caused by PCR-confirmed Plasmodium knowlesi, Plasmodium falciparum, and Plasmodium vivax.
Plasmodium knowlesi can cause severe and fatal human malaria in Southeast Asia. Rapid diagnosis of all Plasmodium species is essential for initiation of effective treatment. Rapid diagnostic tests (RDTs) are sensitive for detection of uncomplicated and severe falciparum malaria but have not been systematically evaluated in knowlesi malaria. At a tertiary referral hospital in Sabah, Malaysia, we prospectively evaluated the sensitivity of two combination RDTs for the diagnosis of uncomplicated and severe malaria from all three potentially fatal Plasmodium species, using a pan-Plasmodium lactate dehydrogenase (pLDH)-P. falciparum histidine-rich protein 2 (PfHRP2) RDT (First Response) and a pan-Plasmodium aldolase-PfHRP2 RDT (ParaHIT). Among 293 hospitalized adults with PCR-confirmed Plasmodium monoinfection, the sensitivity of the pLDH component of the pLDH-PfHRP2 RDT was 74% (95/129; 95% confidence interval [CI], 65 to 80%), 91% (110/121; 95% CI, 84 to 95%), and 95% (41/43; 95% CI, 85 to 99%) for PCR-confirmed P. knowlesi, P. falciparum, and P. vivax infections, respectively, and 88% (30/34; 95% CI, 73 to 95%), 90% (38/42; 95% CI, 78 to 96%), and 100% (12/12; 95% CI, 76 to 100%) among patients tested before antimalarial treatment was begun. Sensitivity in severe malaria was 95% (36/38; 95% CI, 83 to 99), 100% (13/13; 95% CI, 77 to 100), and 100% (7/7; 95% CI, 65 to 100%), respectively. The aldolase component of the aldolase-PfHRP2 RDT performed poorly in all Plasmodium species. The pLDH-based RDT was highly sensitive for the diagnosis of severe malaria from all species; however, neither the pLDH- nor aldolase-based RDT demonstrated sufficiently high overall sensitivity for P. knowlesi. More sensitive RDTs are needed in regions of P. knowlesi endemicity.


Reduced intestinal lipid absorption and body weight-independent improvements in insulin sensitivity in high-fat diet-fed Park2 knockout mice.
Mitochondrial dysfunction is associated with many human diseases and results from mismatch of damage and repair over the life of the organelle. PARK2 is a ubiquitin E3 ligase that regulates mitophagy, a repair mechanism that selectively degrades damaged mitochondria. Deletion of PARK2 in multiple in vivo models results in susceptibility to stress-induced mitochondrial and cellular dysfunction. Surprisingly, Park2 knockout (KO) mice are protected from nutritional stress and do not develop obesity, hepatic steatosis or insulin resistance when fed a high-fat diet (HFD). However, these phenomena are casually related and the physiological basis for this phenotype is unknown. We therefore undertook a series of acute HFD studies to more completely understand the physiology of Park2 KO during nutritional stress. We find that intestinal lipid absorption is impaired in Park2 KO mice as evidenced by increased fecal lipids and reduced plasma triglycerides after intragastric fat challenge. Park2 KO mice developed hepatic steatosis in response to intravenous lipid infusion as well as during incubation of primary hepatocytes with fatty acids, suggesting that hepatic protection from nutritional stress was secondary to changes in energy balance due to altered intestinal triglyceride absorption. Park2 KO mice showed reduced adiposity after 1-wk HFD, as well as improved hepatic and peripheral insulin sensitivity. These studies suggest that changes in intestinal lipid absorption may play a primary role in protection from nutritional stress in Park2 KO mice by preventing HFD-induced weight gain and highlight the need for tissue-specific models to address the role of PARK2 during metabolic stress.


They like to move it (move it): walking kinematics of balitorid loaches of Thailand.
Balitorid loaches are a family of fishes that exhibit morphological adaptations to living in fast flowing water, including an enlarged sacral rib that creates a 'hip'-like skeletal connection between the pelvis and the axial skeleton. The presence of this sacral rib, the robustness of which varies across the family, is hypothesized to facilitate terrestrial locomotion seen in the family. Terrestrial locomotion in balitorids is unlike that of any known fish: the locomotion resembles that of terrestrial tetrapods. Emergence and convergence of terrestrial locomotion from water to land has been studied in fossils; however, studying balitorid walking provides a present-day natural laboratory to examine the convergent evolution of walking movements. We tested the hypothesis that balitorid species with more robust connections between the pelvic and axial skeleton (M3 morphotype) are more effective at walking than species with reduced connectivity (M1 morphotype). We predicted that robust connections would facilitate travel per step and increase mass support during movement. We collected high-speed video of walking in seven balitorid species to analyze kinematic variables. The connection between internal anatomy and locomotion on land are revealed herein with digitized video analysis, μCT scans, and in the context of the phylogenetic history of this family of fishes. Our species sampling covered the extremes of previously identified sacral rib morphotypes, M1 and M3. Although we hypothesized the robustness of the sacral rib to have a strong influence on walking performance, there was not a large reduction in walking ability in the species with the least modified rib (M1). Instead, walking kinematics varied between the two balitorid subfamilies with a generally more 'walk-like' behavior in the Balitorinae and more 'swim-like' behavior in the Homalopteroidinae. The type of terrestrial locomotion displayed in balitorids is unique among living fishes and aids in our understanding of the extent to which a sacral connection facilitates terrestrial walking.


Echo state networks with filter neurons and a delay&sum readout.
Echo state networks (ESNs) are a novel approach to recurrent neural network training with the advantage of a very simple and linear learning algorithm. It has been demonstrated that ESNs outperform other methods on a number of benchmark tasks. Although the approach is appealing, there are still some inherent limitations in the original formulation. Here we suggest two enhancements of this network model. First, the previously proposed idea of filters in neurons is extended to arbitrary infinite impulse response (IIR) filter neurons. This enables such networks to learn multiple attractors and signals at different timescales, which is especially important for modeling real-world time series. Second, a delay&sum readout is introduced, which adds trainable delays in the synaptic connections of output neurons and therefore vastly improves the memory capacity of echo state networks. It is shown in commonly used benchmark tasks and real-world examples, that this new structure is able to significantly outperform standard ESNs and other state-of-the-art models for nonlinear dynamical system modeling.


Long-term nitrogen loading alleviates phosphorus limitation in terrestrial ecosystems.
Increased human-derived nitrogen (N) deposition to terrestrial ecosystems has resulted in widespread phosphorus (P) limitation of net primary productivity. However, it remains unclear if and how N-induced P limitation varies over time. Soil extracellular phosphatases catalyze the hydrolysis of P from soil organic matter, an important adaptive mechanism for ecosystems to cope with N-induced P limitation. Here we show, using a meta-analysis of 140 studies and 668 observations worldwide, that N stimulation of soil phosphatase activity diminishes over time. Whereas short-term N loading (≤5 years) significantly increased soil phosphatase activity by 28%, long-term N loading had no significant effect. Nitrogen loading did not affect soil available P and total P content in either short- or long-term studies. Together, these results suggest that N-induced P limitation in ecosystems is alleviated in the long-term through the initial stimulation of soil phosphatase activity, thereby securing P supply to support plant growth. Our results suggest that increases in terrestrial carbon uptake due to ongoing anthropogenic N loading may be greater than previously thought.


IGF, type I IGF receptor and IGF-binding protein mRNA expression in kidney and liver of potassium-depleted and normal rats infused with IGF-I.
Dietary potassium (K) depletion is known to reduce body weight gain and organ growth, except for kidney which increases in weight. This renal hypertrophy is preceded by increased renal IGF-I levels. In the present study, we investigated IGF-I and -II, type I IGF receptor and IGF-binding protein (IGFBP) mRNA expression in liver and kidney of K-depleted and normal rats infused with vehicle or recombinant human IGF-I. Body weight gain was almost completely arrested in K-depleted rats without any stimulatory effect of IGF-I infusion. Both absolute and relative kidney weight (kidney weight/body weight) were significantly increased in K-depleted rats and this was further enhanced by IGF-I infusion. In contrast, relative liver weight was comparable in the different groups and unaffected by IGF-I infusion. IGF-I mRNA expression was significantly lower in kidney and liver of K-depleted animals whereas type I IGF receptor levels were unchanged. In contrast, in kidney, K depletion increased IGFBP-1 and -2 mRNA expression with no additional effect of IGF-I infusion. In liver of K-depleted animals, IGFBP-1 mRNA expression was increased whereas increased IGFBP-1 and -2 mRNA expression was observed when these animals were infused with IGF-I. These observations may point towards a differential mode of action of the IGFBPs. In kidney increased IGFBP-1 and -2 mRNA expression may enhance IGF-I bioavailability with subsequent kidney growth. In liver, with clearly detectable type I IGF receptor mRNA expression, increased IGFBP levels may protect from IGF-I-induced organ growth by decreasing IGF-I bioavailability.


TigerPlace: training veterinarians about animal companionship for the elderly.
Students learn more effectively when they are actively engaged in the learning process. Therefore, case studies have become increasingly popular as a way to teach students about a representative subject. This article discusses the benefits of case studies, with a primary focus on how case studies can help veterinary medical students learn about the human-animal bond. The discussion is particularly aimed at veterinary medicine instructors and discusses how case studies can be used and why they are important. TigerPlace, a pet-friendly, innovative housing facility for older adults, is used as an example of a case study that can be used to teach about, and to study, the human-animal bond. In particular, the article addresses the special advantages of TigerPlace to students with respect to learning about older adults and the bond they have with their pets.


Computer-Aided Detection (CADx) for Plastic Deformation Fractures in Pediatric Forearm.
Bowing fractures are incomplete fractures of tubular long bones, often observed in pediatric patients, where plain radiographic film is the non-invasive imaging modality of choice in routine radiological workflow. Due to weak association between bent bone and distinct cortex disruption, bowing fractures may not be diagnosed properly while reading plain radiography. Missed fractures and dislocations are common in accidents and emergency practice, particularly in children. These missed injuries can result in more complicated treatment or even long-term disability. The most common reason for missed fractures is that junior radiologists or physicians lack expertise in pediatric skeletal injury diagnosis. Not only is additional radiation exposure inevitable in the case of misdiagnosis, but other consequences include the patient's prolonged uncomfortableness and possible unnecessary surgical procedures. Therefore, a computerized image analysis system, which would be secondary to the radiologists' interpretations, may reduce adverse effects and improve the diagnostic rates of bowing fracture (detection and quantification). This system would be highly desirable and particularly useful in emergency rooms. To address this need, we investigated and developed a new Computer Aided Detection (CADx) system for pediatric bowing fractures. The proposed system has been tested on 226 cases of pediatric forearms with bowing fractures with respect to normal controls. Receiver operation characteristic (ROC) curves show that the sensitivity and selectivity of the developed CADx system are satisfactory and promising. A clinically feasible graphical user interface (GUI) was developed to serve the practical needs in the emergency room as a diagnostic reference. The developed CADx system also has strong potential to train radiology residents for diagnosing pediatric forearm bowing fractures.


Fas signaling induces raft coalescence that is blocked by cholesterol depletion in human RPE cells undergoing apoptosis.
To investigate whether the signaling events occurring in Fas-mediated apoptosis alter raft membrane formation in human RPE cells. Formation of lipid rafts in cultured human retinal pigment epithelial cells (ARPE-19) was studied by confocal microscopy, with fluorescein-labeled cholera toxin subunit B binding protein (BODIPY)-labeled ganglioside GM1 lipid after Fas-L induction of apoptosis. Apoptosis was assessed by fluorescein-labeled annexin V detection of phosphatidylserine externalization and quadrant analysis with flow cytometry. Membrane rafts were localized into membrane vesicles by passing BODIPY-labeled GM1 RPE cells through a 2-microm-pore polycarbonate membrane using an extruder device. The labeled fractions, containing vesicles enriched in GM1, were detected by flow cytometry and then analyzed for the presence of Fas protein. Differential punctate staining of membrane rafts was demonstrated in normal and FasL-induced apoptotic human ARPE-19 cells in culture by confocal microscopy, using cholera toxin B and GM1 labeling of extruded vesicles. The lipid raft-associated vesicles were derived by plasma membrane dissociation, via a newly developed whole-cell extrusion technique that produced 2-microm vesicles with both GM1 lipid and Fas protein abundance enriched in a subpopulation of the membrane-derived vesicles. The amount of Fas protein in the vesicles containing raft domains markedly increased in FasL-treated cells. Treatment of human ARPE 19 cells with methyl beta-cyclodextrin after FasL induction of apoptosis resulted in cellular cholesterol depletion and markedly reduced the incidence of Fas-receptor localization in GM1 rafts. Human ARPE-19 cells in culture contain membrane rafts with apoptotic signaling effectors uniformly distributed in the native state. The cells stimulated to undergo apoptosis appear to use membrane rafts in the death-signaling process by mobilization of rafts to localized regions of the membrane that are now enriched with apoptotic signaling effectors. Fas signaling induces apoptotic raft formation that results in polar condensation, or capping, of the rafts in the late stages of apoptosis. A novel extrusion technique is described that allows localization and enrichment of rafts into membrane vesicles, which can be assayed by flow cytometry. Cholesterol depletion, after Fas ligand activation of apoptosis, reduced raft formation in cells induced to undergo apoptosis. Therapeutic implications for the treatment of retinal disorders are discussed.


Early evidence of effectiveness of digital contact tracing for SARS-CoV-2 in Switzerland.
In the wake of the pandemic of coronavirus disease 2019 (COVID-19), contact tracing has become a key element of strategies to control the spread of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Given the rapid and intense spread of SARS-CoV-2, digital contact tracing has emerged as a potential complementary tool to support containment and mitigation efforts. Early modelling studies highlighted the potential of digital contact tracing to break transmission chains, and Google and Apple subsequently developed the Exposure Notification (EN) framework, making it available to the vast majority of smartphones. A growing number of governments have launched or announced EN-based contact tracing apps, but their effectiveness remains unknown. Here, we report early findings of the digital contact tracing app deployment in Switzerland. We demonstrate proof-of-principle that digital contact tracing reaches exposed contacts, who then test positive for SARS-CoV-2. This indicates that digital contact tracing is an effective complementary tool for controlling the spread of SARS-CoV-2. Continued technical improvement and international compatibility can further increase the efficacy, particularly also across country borders.


Modeling synchronization in networks of delay-coupled fiber ring lasers.
We study the onset of synchronization in a network of N delay-coupled stochastic fiber ring lasers with respect to various parameters when the coupling power is weak. In particular, for groups of three or more ring lasers mutually coupled to a central hub laser, we demonstrate a robust tendency toward out-of-phase (achronal) synchronization between the N-1 outer lasers and the single inner laser. In contrast to the achronal synchronization, we find the outer lasers synchronize with zero-lag (isochronal) with respect to each other, thus forming a set of N-1 coherent fiber lasers.


Indoleamine 2,3-dioxygenase: distribution and function in the developing human placenta.
Studies in mice have suggested that the placenta is protected from immune rejection by maternal T cells by means of localised indoleamine 2,3-dioxygenase dependent depletion of tryptophan. To determine whether such mechanisms might operate in the human placenta, we have studied the physiological importance of human placental indoleamine 2,3-dioxygenase immunohistochemically and functionally. Indoleamine 2,3-dioxygenase is detectable immunohistochemically from day 6 human blastocysts and thereafter throughout pregnancy in syncytiotrophoblasts, extravillous cytotrophoblasts and macrophages in the villous stroma and in the fetal membranes. Interferon-gamma added to villous explants markedly stimulates indoleamine 2,3-dioxygenase protein expression in macrophages. Indoleamine 2,3-dioxygenase-mediated tryptophan degradation in the first trimester villous and decidual tissue explants is stimulated by interferon-gamma and inhibited by 1-methyl-tryptophan (an inhibitor of indoleamine 2,3-dioxygenase). Peripheral blood mononuclear cell proliferation is controlled by indoleamine 2,3-dioxygenase-mediated tryptophan degradation. These results suggest the cellular basis of a mechanism present at the human maternal-fetal interface involved in regulating the maternal immune response to conceptus.


PPARs and adipocyte function.
For long viewed as passive lipid storage depots, adipocytes are now recognised as key players in the pathogenesis of insulin resistance and metabolic disease. In parallel, the last two decades of research have seen the emergence of transcription factors of the peroxisome proliferator-activated receptor (PPAR) family as central regulators of lipid and glucose homeostasis and molecular targets for drugs to treat hyper-lipidaemia and type 2 diabetes mellitus. In this review we discuss the characteristics of PPARs and the role of the different isotypes in adipocyte biology.


A spatially and temporally explicit risk assessment for salmon from a prey base exposed to agricultural insecticides.
This risk assessment applied a framework for determining probable co-occurrence of juvenile spring Chinook salmon (Oncorhynchus tshawytscha) with agricultural pesticides in the Willamette Basin, Oregon (Teply et al. this issue) to characterize risk to the threatened population. The assessment accounted for spatial and temporal distribution of 6 acetylcholinesterase-inhibiting insecticides in salmonid habitat within the basin and their relative contributions to mixture toxicity estimated from chemical monitoring data. The 6 insecticides were chlorpyrifos, diazinon, malathion, carbaryl, carbofuran, and methomyl. Seasonal distributions of the juvenile salmon prey base across the basin were determined and compared to co-occurrence with the insecticide mixture to determine the probability of prey reduction and reduced production of juvenile fish. Probability of effect on freshwater aquatic invertebrates was based on acute toxicity species sensitivity distributions (normalized to the most potent compound, chlorpyrifos) using a novel approach to apply the toxicological concept of concentration addition to species sensitivity distributions with differing slopes. The chlorpyrifos distribution was then used to determine relative sensitivity among various species tested within the important taxa making up the prey base. A prey base index was devised, incorporating diet composition and prey availability, to evaluate the indirect effects of the insecticide mixture on juvenile salmon production occurring as a result of a reduction in the prey base. Our analysis targeted fish use of backwater and off-channel habitat units, because they generally coincide with agricultural lands in lowlands and represent shallow habitat with limited water exchange. The percentage of agricultural land use within 300 m of critical habitat stream reaches was used to scale chemical measurement data from a site with high agricultural land use across the full extent of the basin to provide estimates of chemical exposure in each reach. Seasonal impacts were evaluated from mean monthly concentrations. Stressor impact on 5 key taxa was evaluated at each time step and for each reach, and the outcome was compared to a conservation threshold assigned to the prey base index. Only 13% of juveniles reared in backwater, off-channel habitat within 300 m of agricultural land. Percent reduction of carrying capacity as a consequence of reduced prey was estimated to be 5% over the entire brood year. This can be considered lost capacity that is probably compensated elsewhere via increased occupancy (emigration to other habitat units within the reach), which is not accounted for in the model.


Collectors of natural knowledge: the Edinburgh Medical Society and the associational culture of Scotland and the North Atlantic world in the 18th Century.
This paper reappraises the role of medical clubs and societies in the production and consumption of knowledge in 18th-century Scotland and the wider North Atlantic world. It focuses on the Edinburgh Medical Society, founded in 1731 by Alexander Monro primus; and on the student Medical Society, founded in 1734 and constituted in 1737 as the Medical Society of Edinburgh, ultimately becoming the Royal Medical Society of Edinburgh in 1778. The paper examines how Monro, as editor of the transactions of the Edinburgh Medical Society, sought to adapt medical learning to a world of polite sociability; and how that world came under pressure in the student Medical Society, where prevailing orthodoxies, such as the system of Herman Boerhaave and, later, William Cullen, were challenged. In the febrile atmosphere of the 1790s, William Thomson accused the Royal Medical Society of Edinburgh of promoting visionary theories and abandoning the proper experimental method in medical science. Yet with its overarching commitment to the sceptical and empirical principles laid down by the Royal Society of London (founded in 1660), the Royal Medical Society of Edinburgh provided a model for the establishment of similar clubs and societies on both sides of the Atlantic.


GenomeBlast: a web tool for small genome comparison.
Comparative genomics has become an essential approach for identifying homologous gene candidates and their functions, and for studying genome evolution. There are many tools available for genome comparisons. Unfortunately, most of them are not applicable for the identification of unique genes and the inference of phylogenetic relationships in a given set of genomes. GenomeBlast is a Web tool developed for comparative analysis of multiple small genomes. A new parameter called "coverage" was introduced and used along with sequence identity to evaluate global similarity between genes. With GenomeBlast, the following results can be obtained: (1) unique genes in each genome; (2) homologous gene candidates among compared genomes; (3) 2D plots of homologous gene candidates along the all pairwise genome comparisons; and (4) a table of gene presence/absence information and a genome phylogeny. We demonstrated the functions in GenomeBlast with an example of multiple herpesviral genome analysis and illustrated how GenomeBlast is useful for small genome comparison. We developed a Web tool for comparative analysis of small genomes, which allows the user not only to identify unique genes and homologous gene candidates among multiple genomes, but also to view their graphical distributions on genomes, and to reconstruct genome phylogeny. GenomeBlast runs on a Linux server with 4 CPUs and 4 GB memory. The online version of GenomeBlast is available to public by using a Web browser with the URL http://bioinfo-srv1.awh.unomaha.edu/genomeblast/.


Visual homing using an associative memory.
Homing is the process by which an autonomous system guides itself to a particular location on the basis of sensory input. In this paper, a method of visual homing using an associative memory based on a simple pattern classifier is described. Homing is accomplished without the use of an explicit world model by utilizing direct associations between learned visual patterns and system motor commands. The method is analyzed in terms of a pattern space and conditions obtained that allow the system performance to be predicted on the basis of statistical measurements on the environment. Results of experiments utilizing the method to guide a robot-mounted camera in a three-dimensional environment are presented.


Antibiofilm activity of Actinobacillus pleuropneumoniae serotype 5 capsular polysaccharide.
Cell-free extracts isolated from colony biofilms of Actinobacillus pleuropneumoniae serotype 5 were found to inhibit biofilm formation by Staphylococcus aureus, S. epidermidis and Aggregatibacter actinomycetemcomitans, but not by A. pleuropneumoniae serotype 5 itself, in a 96-well microtiter plate assay. Physical and chemical analyses indicated that the antibiofilm activity in the extract was due to high-molecular-weight polysaccharide. Extracts isolated from a mutant strain deficient in the production of serotype 5 capsular polysaccharide did not exhibit antibiofilm activity. A plasmid harboring the serotype 5 capsule genes restored the antibiofilm activity in the mutant extract. Purified serotype 5 capsular polysaccharide also exhibited antibiofilm activity against S. aureus. A. pleuropneumoniae wild-type extracts did not inhibit S. aureus growth, but did inhibit S. aureus intercellular adhesion and binding of S. aureus cells to stainless steel surfaces. Furthermore, polystyrene surfaces coated with A. pleuropneumoniae wild-type extracts, but not with capsule-mutant extracts, resisted S. aureus biofilm formation. Our findings suggest that the A. pleuropneumoniae serotype 5 capsule inhibits cell-to-cell and cell-to-surface interactions of other bacteria. A. pleuropneumoniae serotype 5 capsular polysaccharide is one of a growing number of bacterial polysaccharides that exhibit broad-spectrum, nonbiocidal antibiofilm activity. Future studies on these antibiofilm polysaccharides may uncover novel functions for bacterial polysaccharides in nature, and may lead to the development of new classes of antibiofilm agents for industrial and clinical applications.


The host STING pathway at the interface of cancer and immunity.
A major subset of human cancers shows evidence for spontaneous adaptive immunity, which is reflected by the presence of infiltrating CD8+ T cells specific for tumor antigens within the tumor microenvironment. This observation has raised the question of which innate immune sensing pathway might detect the presence of cancer and lead to a natural adaptive antitumor immune response in the absence of exogenous infectious pathogens. Evidence for a critical functional role for type I IFNs led to interrogation of candidate innate immune sensing pathways that might be triggered by tumor presence and induce type I IFN production. Such analyses have revealed a major role for the stimulator of IFN genes pathway (STING pathway), which senses cytosolic tumor-derived DNA within the cytosol of tumor-infiltrating DCs. Activation of this pathway is correlated with IFN-β production and induction of antitumor T cells. Based on the biology of this natural immune response, pharmacologic agonists of the STING pathway are being developed to augment and optimize STING activation as a cancer therapy. Intratumoral administration of STING agonists results in remarkable therapeutic activity in mouse models, and STING agonists are being carried forward into phase I clinical testing.


H2, HD, and D2 inside C60: coupled translation-rotation eigenstates of the endohedral molecules from quantum five-dimensional calculations.
We have performed rigorous quantum five-dimensional (5D) calculations of the translation-rotation (T-R) energy levels and wave functions of H(2), HD, and D(2) inside C(60). This work is an extension of our earlier investigation of the quantum T-R dynamics of H(2)@C(60) [M. Xu et al., J. Chem. Phys. 128, 011101 (2008)] and uses the same computational methodology. Two 5D intermolecular potential energy surfaces (PESs) were employed, differing considerably in their well depths and the degree of confinement of the hydrogen molecule. Our calculations revealed pronounced sensitivity of the endohedral T-R dynamics to the differences in the interaction potentials, and to the large variations in the masses and the rotational constants of H(2), HD, and D(2). The T-R levels vary significantly in their energies and ordering on the two PESs, as well as from one isotopomer to another. Nevertheless, they all display the same distinctive patterns of degeneracies, which can be qualitatively understood and assigned in terms the model which combines the isotropic three-dimensional harmonic oscillator, the rigid rotor, and the coupling between the orbital and the rotational angular momenta of H(2)/HD/D(2). The quantum number j associated with the rotation of H(2), HD, and D(2) was found to be a good quantum number for H(2) and D(2) on both PESs, while most of the T-R levels of HD exhibit strong mixing of two or more rotational basis functions with different j values.


Streptococcus pneumoniae and noncapsular Haemophilus influenzae nasal carriage and hand contamination in children: a comparison of two populations at risk of otitis media.
Australian Indigenous children living in remote areas have rates of tympanic membrane perforation as high as 60%, almost 100 times the prevalence in urban child care settings (<1%). Relative rates of pneumococcal nasal carriage do not reflect this difference in disease risk. Cross-sectional comparison of nasal carriage and hand contamination in children younger than 4 years of age from urban child-care centers and Indigenous children 3-7 years of age from a remote community. Almost identical methods of nasal swab collection, transport and culture were used. Data on pneumococcal antimicrobial susceptibility patterns and serotypes are also reported. For Indigenous children compared with children in child care, the relative risk of nasal carriage of either pneumococcus or noncapsular Haemophilus influenzae was <2-fold [relative risk, 1.7; 95% confidence interval (CI), 1.5, 1.9], the risk of simultaneous nasal carriage was almost 3-fold (78% versus 28%; relative risk, 2.9; 95% CI 2.3, 3.5), and the risk of pneumococcal hand contamination was 8-fold higher (37% versus 4%; relative risk, 8.4; 95% CI 4.6, 15.2). For simultaneous hand contamination, the risk was 23-fold (8% versus 0.3%; relative risk, 23.1; 95% CI 2.9, 185.4). Remote Indigenous children also had a more diverse serotype distribution (25 versus 14 serotypes identified). Simultaneous nasal carriage of Streptococcus pneumoniae and H. influenzae and hand contamination are simple indicators of risk for use in studies of otitis media in populations at risk for tympanic membrane perforation.


Highly stabilized lipase in polyaniline nanofibers for surfactant-mediated esterification of ibuprofen.
Lipase (LP) from Candida rugosa was immobilized and stabilized in polyaniline nanofibers (PANFs) via a three-step process of enzyme adsorption, precipitation, and cross-linking, which generates the final immobilization called "EAPC". The activity of EAPC was 5.1 and 5.9 times higher than those of LP immobilizations via enzyme adsorption (EA) and enzyme adsorption/cross-linking (EAC), respectively. After incubation in an aqueous buffer under shaking (200 rpm) for 84 days, EAPC maintained 74% of its initial activity, while EA and EAC retained 11 and 24% of their initial activities, respectively. Highly stable and active EAPC was employed for the resolution of racemic ibuprofen via esterification of S-(+)-ibuprofen with 1-propanol in isooctane. The addition of 100 mM dioctyl sulfosuccinate (AOT) into the reaction medium increased the esterification activity by 61-fold, which can be explained by the better dispersion of EAPC in isooctane. EAPC showed 42% conversion in the esterification of racemic ibuprofen after 102 h, whereas EA and EAC showed only 1.2 and 1.4% conversion in the same condition, respectively. The EAPC approach increases both loading and stability of LP, and the combination of EAPC with the surfactant addition can be employed for efficient enzymatic reactions in organic solvents.


Retinoic acid induces homing of protective T and B cells to the gut after subcutaneous immunization in mice.
Diarrheal diseases represent a major health burden in developing countries. Parenteral immunization typically does not induce efficient protection against enteropathogens because it does not stimulate migration of immune cells to the gut. Retinoic acid (RA) is critical for gut immunity, inducing upregulation of gut-homing receptors on activated T cells. In this study, we have demonstrated that RA can redirect immune responses elicited by s.c. vaccination of mice from skin-draining inguinal LNs (ingLNs) to the gut. When present during priming, RA induced robust upregulation of gut-homing receptors in ingLNs, imprinting gut-homing capacity on T cells. Concurrently, RA triggered the generation of gut-tropic IgA+ plasma cells in ingLNs and raised the levels of antigen-specific IgA in the intestinal lumen and blood. RA applied s.c. in vivo induced autonomous RA production in ingLN DCs, further driving efficient induction of gut-homing molecules on effector cells. Importantly, RA-supplemented s.c. immunization elicited a potent immune response in the small intestine that protected mice from cholera toxin–induced diarrhea and diminished bacterial loads in Peyer patches after oral infection with Salmonella. Thus, the use of RA as a gut-homing navigator represents a powerful tool to induce protective immunity in the intestine after s.c. immunization, offering what we believe to be a novel approach for vaccination against enteropathogens.


Improved Dermal Regeneration Using a Combination of Dermal Substitutes and Dermal Fibroblast Optimization: A Hypothesis.
In human adults, the repair of cutaneous wounds usually leads to scar formation rather than regeneration. Dermal substitutes have been used as a regenerative template for reducing scar formation and improving the extent of dermal regeneration. However, achievement of complete regeneration is still a long way off. Dermal substitutes are characterized by unusual regenerative activity, appearing to function by acting as temporary configurational guides for cell infiltration and synthesis of new stroma. Fibroblasts are important cells with many vital functions in wound-healing processes. They are heterogeneous with distinct characteristics according to their source location, such as subcutaneous tissue, superficial-layer dermis, and deep-layer dermis. Many studies have shown that superficial dermal fibroblasts possess the potential to form dermis-like tissue. Fibroblasts in deep-layer dermis and subcutaneous tissue may play a critical role in the formation of hypertrophic scars. Fibroblast phenotype affects the newly formed dermal architecture and influences the dermal regeneration effect induced by dermal substitutes. It is hypothesized that better regeneration of the dermis can be achieved using dermal substitutes along with dermal fibroblast optimization.


Smoking cessation support delivered via mobile phone text messaging (txt2stop): a single-blind, randomised trial.
Smoking cessation programmes delivered via mobile phone text messaging show increases in self-reported quitting in the short term. We assessed the effect of an automated smoking cessation programme delivered via mobile phone text messaging on continuous abstinence, which was biochemically verified at 6 months. In this single-blind, randomised trial, undertaken in the UK, smokers willing to make a quit attempt were randomly allocated, using an independent telephone randomisation system, to a mobile phone text messaging smoking cessation programme (txt2stop), comprising motivational messages and behavioural-change support, or to a control group that received text messages unrelated to quitting. The system automatically generated intervention or control group texts according to the allocation. Outcome assessors were masked to treatment allocation. The primary outcome was self-reported continuous smoking abstinence, biochemically verified at 6 months. All analyses were by intention to treat. This study is registered, number ISRCTN 80978588. We assessed 11,914 participants for eligibility. 5800 participants were randomised, of whom 2915 smokers were allocated to the txt2stop intervention and 2885 were allocated to the control group; eight were excluded because they were randomised more than once. Primary outcome data were available for 5524 (95%) participants. Biochemically verified continuous abstinence at 6 months was significantly increased in the txt2stop group (10·7% txt2stop vs 4·9% control, relative risk [RR] 2·20, 95% CI 1·80-2·68; p<0·0001). Similar results were obtained when participants that were lost to follow-up were treated as smokers (268 [9%] of 2911 txt2stop vs 124 [4%] of 2881 control [RR 2·14, 95% CI 1·74-2·63; p<0·0001]), and when they were excluded (268 [10%] of 2735 txt2stop vs 124 [4%] of 2789 control [2·20, 1·79-2·71; p<0·0001]). No significant heterogeneity was shown in any of the prespecified subgroups. The txt2stop smoking cessation programme significantly improved smoking cessation rates at 6 months and should be considered for inclusion in smoking cessation services. UK Medical Research Council, Primary Care Research Networks.


Association between different hemoglobin A1c levels and clinical outcomes among elderly nursing home residents with type 2 diabetes mellitus.
New guidelines recommend a target glycated hemoglobin (HbA1c) of 7.5% to 8.0% in elderly persons with type 2 diabetes mellitus (T2DM), but real-world data regarding outcomes associated with different HbA1c levels in the elderly are limited. This study assessed outcomes and their association with defined HbA1c thresholds and age ranges in insulin-treated, elderly, patients with T2DM in long-term care (LTC). Retrospective analysis of medical charts and the Minimum Data Set (MDS) for the period September 2010 through September 2011. A total of 117 nursing homes in the United States. Eligible patients had resided in LTC for 3 months or more, had at least 1 full MDS assessment, 2 or more records of insulin dispensing with no pump use, and 1 or more HbA1c measurements. Outcomes that were measured included hypoglycemia, ketoacidosis, infections, falls, hospitalization, and emergency room (ER) visits. A total of 583 patients were included (mean age 78.9 years, mean chart observation length 55 days). In all groups, hypoglycemia was lowest in patients with an HbA1c level higher than 9.0%. In patients 75 years or older, infection rates were highest when HbA1c levels were higher than 9.0%. Falls increased by HbA1c level in patients aged 65 to 74 years, but decreased by HbA1c levels in patients 85 years or older. Ketoacidosis, hospitalization, and ER visits were low in all groups. These data suggest that better glycemic levels may not necessarily be associated with better clinical outcomes, and different age groups may exhibit different patterns, thereby supporting the call for individualized glycemic control among elderly patients.


Bismuth coating of non-tunneled haemodialysis catheters reduces bacterial colonization: a randomized controlled trial.
Haemodialysis (HD) catheter-related blood stream infections are a major cause of morbidity and mortality in patients with acute and chronic renal failure. We conducted a randomized, prospective, double-blinded trial investigating the clinical value of bismuth-coated non-tunneled HD catheters in patients in need of temporary short-term vascular access. A standard catheter (SC) was compared to a surface-modified, bismuth-film-coated catheter (FCC). After removal of the catheter for any reason, both arterial and venous lumina were rinsed and the fluid cultured for detection of bacterial colony-forming units (CFU). The catheter tip was placed in a tube containing sterile saline, sonicated and shortly centrifuged to remove debris (3 min at 1000 g). The supernatant was cultured and assayed for DNA content. Seventy-seven patients in three HD units were randomized. Thirteen patients suffered from acute renal failure, 60 patients from chronic renal failure, and four patients without renal insufficiency were treated with plasma exchange. The time to catheter removal was not significantly different between groups, with a mean of 18.5 +/- 2 days for SC and 15.1 +/- 2 days for FCC. In most cases, the reasons for catheter removal were related to no further need for extracorporeal therapy or establishment of a permanent vascular access. Six catheters for SC and four catheters for FCC were removed because of presumed infection. Bacterial colonization was significantly lower for coated catheters compared to standard catheters, both for cultured catheter tips as well as for CFU in rinse fluids (P < 0.05). Surface modification with bismuth film reduces bacterial colonization of temporary non-tunneled HD catheters in a clinical trial. Larger trials with these modified catheters are justified to further investigate the effect on catheter-related infections, complications and costs.


Prevalence and correlates of doctor-geriatric patient lifestyle discussions: analysis of ADEPT videotapes.
This study identifies the prevalence and correlates of physician-geriatric patient discussions about physical activity and nutrition lifestyle behaviors. Between August 1998 and July 2000, 423 older patient visits to 36 physicians were videotaped in three different primary care settings. The patient sample was primarily white, female, well-educated, and financially sufficient, although 12.7% of the encounters occurred in an inner city clinic. The major dependent variable-lifestyle discussion-is based on observations of physician behavior. Descriptive and multivariate logistic regression analyses were conducted in 2004. Nutrition talk was most prevalent, occurring in almost half the encounters (48.2%) followed by physical activity discussions (39.2%) then conjoint mention (22%). Discussions were significantly less likely to occur in acute visits. While ethnicity, gender, and length of visit were not significantly related, physician interaction style and patient vitality and education were significant predictors in the multivariate analyses. Given the impact of lifestyle behaviors on myriad health outcomes, the current prevalence rates of physician discussion, while higher than in many previous studies, remain sub-optimal. Practical assessment tools, training in behavioral counseling, and reimbursement incentives are recommended strategies for raising physical activity and nutrition discussion prevalence in primary care settings.


The utility of CA 19-9 in the diagnoses of cholangiocarcinoma in patients without primary sclerosing cholangitis.
The diagnosis of cholangiocarcinoma is often difficult, making management approaches problematic. A reliable serum tumor marker for cholangiocarcinoma would be a useful additional diagnostic test. Previous studies have demonstrated that elevated serum concentrations of CA 19-9, a tumor-associated antigen, have good sensitivity and specificity for cholangiocarcinoma in patients with primary sclerosing cholangitis. However, the value of this tumor marker for cholangiocarcinoma unassociated with primary sclerosing cholangitis is unclear. Thus, the aims of this study were to determine the usefulness of a serum CA 19-9 determination in the diagnosis of de novo cholangiocarcinoma. We prospectively measured serum CA 19-9 concentrations in patients with cholangiocarcinoma (n = 36), nonmalignant liver disease (n = 41), and benign bile duct strictures (n = 26). Serum CA 19-9 concentrations were measured by an immunoradiometric assay (CIS Bio International) without knowledge of the clinical diagnosis. The sensitivity of a CA 19-9 value >100 U/ml in diagnosing cholangiocarcinoma was 53%. When compared with the nonmalignant liver disease and the benign bile duct stricture groups, the true negative rates were 76% and 92%, respectively. Patients with unresectable cholangiocarcinoma had significantly greater mean CA 19-9 concentrations compared to patients with resectable cholangiocarcinoma. These data suggest that the serum CA 19-9 determination is a useful addition to the available tests for the differential diagnosis of cholangiocarcinoma.


Sex and acute weighted vest differences in force production and joint work during countermovement vertical jumping.
It is unclear whether weighted vest (WV) use improves countermovement vertical jump (CMVJ) performance by enhancing stretch-shortening cycle (SSC) function via increased storage and utilisation of elastic strain energy. In is also unknown whether WV use stimulates different responses in men and women. WV effects on energy storage and utilisation during CMVJ were examined in men and women. Nine men (25 ± 3 y; 89.7 ± 18.7 kg; 1.8 ± 0.1 m) and 12 women (24 ± 3 y; 62.7 ± 10.3 kg; 1.6 ± 0.1 m) performed CMVJ wearing a WV with (loaded) and without (unloaded) 10% added mass while kinematic and ground reaction force (GRF) data were obtained. A longer eccentric sub-phase and increased storage of elastic strain energy occurred when loaded. Increased positive joint work occurred during the concentric portion of loaded CMVJ. Women exhibited less positive hip work and greater positive ankle work than men during the unloading and eccentric sub-phases, respectively. Joint work was similar between sexes during the concentric sub-phase, likely due to decreased trunk extension excursion in men when loaded. Women and men employ different SSC strategies during the CMVJ, though the different strategies do not alter energy storage or concentric mechanical output.


Entropy: a conceptual approach to measuring situation-level workload within emergency care and its relationship to emergency department crowding.
Emergency department (ED) crowding correlates with patient safety. Difficulties quantifying crowding and providing solutions were highlighted in the recent Institute of Medicine (IOM) report calling for the application of advanced industrial engineering (IE) research techniques to evaluate ED crowding. ED personnel workload is a related concept, with potential reciprocal effects between the two. Collaboration between emergency medicine and IE is needed to address crowding and ED personnel workload. We review ED crowding and workload literature, relationships between workload and ED crowding, and the potential application of information theory as implemented in IE frameworks entitled "entropy" in evaluating both topics. IE techniques have applications for emergency medicine and have been successful in helping improve ED operations. Lean and Six Sigma applications are some of these techniques. Existing ED workload measures don't account for all aspects of work in the ED (acuity, efficiency, tasks, etc.) Crowding scales, such as NEDOCS (National ED Overcrowding Study) and EDWIN (ED Work Index), fail to predict ED crowding. A new measurement "entropy" may provide a more comprehensive evaluation of ED workload and may predict work overload seen with crowding. Entropy measures task-based work and the information flow involved. By assigning an entropy value to patient type-specific tasks, we might predict when the ED is overwhelmed, and crowded. IE techniques provide solutions to the ED crowding problem and improve ED workload. We propose a technique novel to medicine: "Entropy," derived from information theory, which may provide insight into ED personnel workload, its potential for measuring ED crowding, and possibly, in predicting an overwhelming situation.


Alternatives to placebo-controlled trials.
Until recently, the gold standard for assessing the efficacy and effectiveness of new medications has been the placebo-control randomized clinical trial (RCT). However, there are serious ethical concerns about placing patients on a placebo when effective treatments exist. Further, if a new agent is tested only against a placebo, there is no guarantee that it is more effective, or even as effective, as an existing agent. For these and other reasons, ethicists and regulatory bodies have said that, under these circumstances, new drugs should be tested against an active agent. There are three types of such trials: superiority, equivalence, and non-inferiority. In superiority trials, the goal is to establish that the new drug is better (i.e., more effective, or with a more benign side-effect profile) than the standard. Because such trials require much larger sample sizes than placebo-control studies, and are rarely required to bring a drug onto market, they are rarely done. In equivalence trials, the aim is to show that the new and standard agents have similar degrees of effectiveness or adverse events. Due to sample size requirements, most studies of new drugs are non-inferiority trials, in which it is sufficient to demonstrate that the new drug is not significantly worse than the existing ones. However, there are methodological concerns with equivalence and non-inferiority trials, including (a) an inability to determine if the drugs were equally good or equally bad; (b) poorly executed trials with low power can be mistaken for "proving" equivalence or non-inferiority; (c) the equivalence interval is arbitrary; (d) successive non-inferiority trials may lead to a gradual reduction in effectiveness; and (e) often larger trials are necessary. The paper also discusses "add on trials." It is recommended that, even when existing drugs exist, trials should consist of at least three arms, one of which is a placebo. This paper briefly considers the ethics of placebo, and conditions are stated under which such studies can be conducted.


When cheating would make you a cheater: implicating the self prevents unethical behavior.
In 3 experiments using 2 different paradigms, people were less likely to cheat for personal gain when a subtle change in phrasing framed such behavior as diagnostic of an undesirable identity. Participants were given the opportunity to claim money they were not entitled to at the experimenters' expense; instructions referred to cheating with either language that was designed to highlight the implications of cheating for the actor's identity (e.g., "Please don't be a cheater") or language that focused on the action (e.g., "Please don't cheat"). Participants in the "cheating" condition claimed significantly more money than did participants in the "cheater" condition, who showed no evidence of having cheated at all. This difference occurred both in a face-to-face interaction (Experiment 1) and in a private online setting (Experiments 2 and 3). These results demonstrate the power of a subtle linguistic difference to prevent even private unethical behavior by invoking people's desire to maintain a self-image as good and honest.


Invasion success in a marginal habitat: an experimental test of competitive ability and drought tolerance in Chromolaena odorata.
Climatic niche models based on native-range climatic data accurately predict invasive-range distributions in the majority of species. However, these models often do not account for ecological and evolutionary processes, which limit the ability to predict future range expansion. This might be particularly problematic in the case of invaders that occupy environments that would be considered marginal relative to the climatic niche in the native range of the species. Here, we assess the potential for future range expansion in the shrub Chromolaena odorata that is currently invading mesic savannas (>650 mm MAP) in South Africa that are colder and drier than most habitats in its native range. In a greenhouse experiment we tested whether its current distribution in South Africa can be explained by increased competitive ability and/or differentiation in drought tolerance relative to the native population. We compared aboveground biomass, biomass allocation, water use efficiency and relative yields of native and invasive C. odorata and the resident grass Panicum maximum in wet and dry conditions. Surprisingly, we found little differentiation between ranges. Invasive C. odorata showed no increased competitive ability or superior drought tolerance compared to native C. odorata. Moreover we found that P. maximum was a better competitor than either native or invasive C. odorata. These results imply that C. odorata is unlikely to expand its future range towards more extreme, drier, habitats beyond the limits of its current climatic niche and that the species' invasiveness most likely depends on superior light interception when temporarily released from competition by disturbance. Our study highlights the fact that species can successfully invade habitats that are at the extreme end of their ranges and thereby contributes towards a better understanding of range expansion during species invasions.


Tuning the redox profiles of polyoxovanadate-alkoxide clusters via heterometal installation: toward designer redox Reagents.
Herein, we describe a rational synthetic approach for tuning the electrochemical profiles of a series of Lindqvist polyoxovanadate-alkoxide clusters through heterometal functionalization. Synthetic procedures for group(IV) functionalization of the mixed-valent POV-alkoxide cluster, [V6O7(OCH3)12], are established, resulting in the heterometallic species, [NBu4][V5O6(OCH3)12MOCH3] (M = Ti, Zr, Hf). We demonstrate that these d0, heterometallic dopants anodically shift the potential of the electrochemical processes associated with the cluster, making the molecule more resistant to oxidation. Conversely, incorporation of electron rich heterometals yields a more readily oxidized molecule, with redox processes shifted cathodically. The predictable tuning and remarkable electrochemical profiles of this family of heterometal-functionalized polyoxometalates highlights their potential use as designer redox agents.


Experiences and consequences of pain in persons with post-polio syndrome.
This study describes the meaning of pain and its implications for everyday life in 35 persons with symptoms of post-polio syndrome. The mean age of the study group is 65 years and the sex ratio of men to women is 1.5:1. The study persons were interviewed on two occasions in their homes and answered a pain questionnaire. The result shows that everyday vocabulary is used to express pain experiences. The study persons normally answered that it hurt, although the interviewers used pain in their questions. The results show that the lower back is the most common location of pain. Joint pains are most common in the upper extremities. The pain is worst in the evening and at night, and tangibly affects the daily rhythm. Physical strain and climatic factors commonly provoke pain, whereas rest and heat give relief. The study show that interviews and pain questionnaire should be supplemented with questions on activities so as to gain a comprehensive view of the difficulties experienced in everyday life.


Effect of postdiagnosis weight change on hot flash status among early-stage breast cancer survivors.
Hot flashes (HF) affect a large proportion of breast cancer (BC) survivors and can negatively affect their quality of life. Treatments other than estrogen replacement to alleviate HF are needed. Body weight is related to hot flashes, but little is known about the effect of weight change on HF. We used data from 3,088 women previously treated for early-stage BC who were enrolled onto the Women's Healthy Eating and Living study to examine the association between weight change after a breast cancer diagnosis and the odds of reporting HF. Overall, 36.1% of participants reported moderate to severe HF at study entry. At 2 years postdiagnosis, 69.2% of women remained within 10%, 4.8% lost at least 10%, and 26.0% gained at least 10% of their prediagnosis weight. Those who gained at least 10% of their prediagnosis weight had a greater risk of reporting HF than women who remained weight stable in that same period (odds ratio [OR], 1.33; 95% CI, 1.11 to 1.60; P = .003). Weight loss of at least 10% of prediagnosis weight was associated with a nonsignificant reduced risk (OR, 0.72; 95% CI, 0.47 to 1.08; P = .118) of reporting HF. However, the trend of weight change (weight loss and weight gain) on HF was significant both when examined categorically (P = .03) and continuously (P < .001). Prevention of weight gain after a BC diagnosis-a modifiable behavior-may offer a viable intervention for relief of HF. Effects of intentional weight loss in BC survivors requires further study.


Outcome of Flow Diverters with Surface Modifications in Treatment of Cerebral Aneurysms: Systematic Review and Meta-analysis.
Newer flow diverters are enhanced with antithrombogenic surface modifications like the Pipeline Embolization Device with Shield Technology and the Derivo Embolization Device and are purported to facilitate deployment and reduce ischemic events. Our aim was to review the safety and efficacy of surface-modified flow diverters in treating patients with cerebral aneurysms. We used Preferred Reporting Items for Systematic Reviews and Meta-Analyses-compliant systematic review and meta-analysis covering 3 major data bases and gray literature between 2014 and 2019. Two reviewers independently reviewed human studies of surface-modified flow diverters for eligibility based on predetermined criteria. The random effects model and Freeman-Tukey arcsine transformation were used to pool efficacy outcomes (technical success, aneurysm occlusion at 6 and 12 months) and safety outcomes (mortality, morbidity, all ischemia, and serious ischemia). Subgroup analysis was performed to compare outcomes between 2 different flow diverters. Eight single-arm case series involving 911 patients and 1060 aneurysms were included. The median follow-up was 8.24 months. Pooled estimate for technical success was 99.6%, while the aneurysm occlusion at 6 and 12 months were 80.5%, and 85.6%, respectively. Pooled estimates for mortality, morbidity, total ischemia, and serious ischemia rates were 0.7%, 6.0%, 6.7%, and 1.8%, respectively. Most studies were of good quality, and no significant heterogeneity was observed. Limitations include a retrospective, observational design in some studies; heterogeneous and underreported antiplatelet therapy; and potential performance and ecologic bias. Early-to-midterm safety and efficacy for surface-modified flow diverters appear comparable with older devices, especially for small, unruptured anterior circulation aneurysms. Long-term clinical data are required to further corroborate these results.


Internet-based treatment for adults with depressive symptoms: the protocol of a randomized controlled trial.
Depression is a highly prevalent condition, affecting more than 15% of the adult population at least once in their lives. Guided self-help is effective in the treatment of depression. The purpose of this study is to investigate the effectiveness of two Internet-based guided self-help treatments with adults reporting elevated depressive symptoms. Other research questions concern the identification of potential mediators and the search for subgroups who respond differently to the interventions. This study is a randomized controlled trial with three conditions: two treatment conditions and one waiting list control group. The two treatment conditions are Internet-based cognitive behavior therapy and Internet-based problem-solving therapy. They consist of 8 and 5 weekly lessons respectively. Both interventions are combined with support by e-mail. Participants in the waiting list control group receive the intervention three months later. The study population consists of adults from the general population. They are recruited through advertisements in local and national newspapers and through banners on the Internet. Subjects with symptoms of depression (> or = 16 on the Center for Epidemiological Studies Depression scale) are included. Other inclusion criteria are having sufficient knowledge of the Dutch language, access to the Internet and an e-mail address. Primary outcome is depressive symptoms. Secondary outcomes are anxiety, quality of life, dysfunctional cognitions, worrying, problem solving skills, mastery, absence at work and use of healthcare. We will examine the following variables as potential mediators: dysfunctional cognitions, problem solving skills, worrying, anxiety and mastery. Potential moderating variables are: socio-demographic characteristics and symptom severity. Data are collected at baseline and at 5 weeks, 8 weeks, 12 weeks and 9 months after baseline. Analyses will be conducted on the intention-to-treat sample. This study evaluates two Internet-based treatments for depression, namely cognitive behavioral therapy and problem-solving therapy. The effectiveness of Internet-based problem-solving therapy suggest that this may be a worthwhile alternative to other more intensive treatment options. Strengths and limitations of this study are discussed. Current Controlled Trials ISRCTN16823487.


Modular capsid decoration boosts adenovirus vaccine-induced humoral immunity against SARS-CoV-2.
Adenovirus vector vaccines have been widely and successfully deployed in response to coronavirus disease 2019 (COVID-19). However, despite inducing potent T cell immunity, improvement of vaccine-specific antibody responses upon homologous boosting is modest compared with other technologies. Here, we describe a system enabling modular decoration of adenovirus capsid surfaces with antigens and demonstrate potent induction of humoral immunity against these displayed antigens. Ligand attachment via a covalent bond was achieved using a protein superglue, DogTag/DogCatcher (similar to SpyTag/SpyCatcher), in a rapid and spontaneous reaction requiring only co-incubation of ligand and vector components. DogTag was inserted into surface-exposed loops in the adenovirus hexon protein to allow attachment of DogCatcher-fused ligands on virus particles. Efficient coverage of the capsid surface was achieved using various ligands, with vector infectivity retained in each case. Capsid decoration shielded particles from vector neutralizing antibodies. In prime-boost regimens, adenovirus vectors decorated with the receptor-binding domain of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) spike induced >10-fold higher SARS-CoV-2 neutralization titers compared with an undecorated vector encoding spike. Importantly, decorated vectors achieved equivalent or superior T cell immunogenicity against encoded antigens compared with undecorated vectors. We propose capsid decoration using protein superglues as a novel strategy to improve efficacy and boostability of adenovirus-based vaccines and therapeutics.


Inhibition of intraerythrocytic development of Plasmodium falciparum by proteinase inhibitors.
A group of inactivators of cysteinyl proteinases which function by covalent bond formation have been examined for their ability to inhibit the development of Plasmodium falciparum within red blood cells. The most effective of these caused inactivation of the parasite near 10(-8) M concentration. The range of inhibitory action varied with peptide structure in a manner characteristic of affinity labels for proteinases suggesting that the target of inhibition was an unidentified proteinase, probably of the cysteinyl type, but different from cathepsins B and L.


HLA-G remains a mystery.
In this brief summary, we argue that many widely held beliefs about HLA-G are questionable. Recent research has led to a re-evaluation of many of the characteristics that were thought to make HLA-G unusual among the MHC class I molecules. First, contrary to reports suggesting that the gene encoding HLA-G exhibits marked polymorphism in some human populations, recent data have shown that the HLA-G gene has comparatively little polymorphism - a feature that might allow it to be expressed in the placenta without causing rejection by the maternal immune system. Second, although truncated forms of HLA-G are generated in the placenta, most of them are unlikely to have significant biological effects as they do not reach the cell surface. Third, the hypothesis that a major role of HLA-G is to prevent attack of the placenta by maternal natural killer cells is now the subject of renewed scrutiny. Finally, there is little evidence that the induction of expression of HLA-G is a major mechanism by which tumor cells avoid immune attack. HLA-G has once again become as mysterious as when it was discovered: an MHC class I molecule expressed at a challengingly extraordinary site--the immunologically uneasy interface between mother and fetus.


A key to new world distatrix mason (Hymenoptera: Braconidae), with descriptions of six new reared neotropical species.
Six new species of the genus Distatrix Mason from Central and South America, D. loretta, D. xanadon, D. vigilis, D. pitillaensis, D. pandora Grinter, n. sp., and D. antirrheae Whitfield & Grinter, n. sp., are described from large-scale caterpillar inventory endeavors, mostly from the larvae of geometrid moths. Biological information and diagnostic features that distinguish these species from other previously described Distatrix, especially those from the Neotropical region, are provided; and the first key to New World species is presented. The new discoveries expand our knowledge of the World's widespread Distatrix fauna by about a third, suggesting that similar survey efforts in other poorly sampled regions will reveal numerous additional undescribed species.


Identifying the emergence of the superficial peroneal nerve through deep fascia on ultrasound and by dissection: Implications for regional anesthesia in foot and ankle surgery.
Regional anesthesia relies on a sound understanding of anatomy and the utility of ultrasound in identifying relevant structures. We assessed the ability to identify the point at which the superficial peroneal nerve (SPN) emerges through the deep fascia by ultrasound on 26 volunteers (mean age 27.85 years ± 13.186; equal male: female). This point was identified, characterized in relation to surrounding bony landmarks (lateral malleolus and head of the fibula), and compared to data from 16 formalin-fixed human cadavers (mean age 82.88 years ± 6.964; equal male: female). The SPN was identified bilaterally in all subjects. On ultrasound it was found to pierce the deep fascia of the leg at a point 0.31 (±0.066) of the way along a straight line from the lateral malleolus to the head of the fibula (LM-HF line). This occurred on or anterior to the line in all cases. Dissection of cadavers found this point to be 0.30 (±0.062) along the LM-HF line, with no statistically significant difference between the two groups (U = 764.000; exact two-tailed P = 0.534). It was always on or anterior to the LM-HF line, anterior by 0.74 cm (±0.624) on ultrasound and by 1.51 cm (±0.509) during dissection. This point was significantly further anterior to the LM-HF line in cadavers (U = 257.700, exact two-tailed P < 0.001). Dissection revealed the nerve to divide prior to emergence in 46.88% (n = 15) limbs, which was not identified on ultrasound (although not specifically assessed). Such information can guide clinicians when patient factors (e.g., obesity and peripheral edema) make ultrasound-guided nerve localization more technically challenging. Clin. Anat. 32:390-395, 2019. © 2019 Wiley Periodicals, Inc.


Synthesis, in vitro antiviral evaluation, and stability studies of novel alpha-borano-nucleotide analogues of 9-[2-(phosphonomethoxy)ethyl]adenine and (R)-9-[2-(phosphonomethoxy)propyl]adenine.
We describe here the synthesis of 9-[2-(boranophosphonomethoxy)ethyl]adenine (6a) and (R)-9-[2-(boranophosphonomethoxy)propyl]adenine (6b), the first alpha-boranophosphonate nucleosides in which a borane (BH3) group substitutes one nonbridging oxygen atom of the alpha-phosphonate moiety. H-phosphinates 5a and 5b and alpha-boranophosphonates 6a and 6b were evaluated for their in vitro activity against human immunodeficiency virus (HIV) infected cells and against a panel of DNA or RNA viruses. Compounds 5a, 5b, 6a, and 6b exhibited no significant antiviral activity in vitro and cytotoxicity. To measure the chemical and enzymatic stabilities of the target compounds 6a and 6b, kinetic data of decomposition for derivatives 5a, 5b, 6a, 6b, and standard compounds were studied at 37 degrees C in several media. The alpha-boranophosphonates 6a and 6b were metabolized in culture medium into H-phosphinates 5a and 5b, with half-live values of 5.3 h for 6a and 1.3 h for 6b.


The role of serotonin in personality inference: tryptophan depletion impairs the identification of neuroticism in the face.
Serotonergic mechanisms mediate the expression of personality traits (such as impulsivity, aggression and anxiety) that are linked to vulnerability to psychological illnesses, and modulate the identification of emotional expressions in the face as well as learning about broader classes of appetitive and aversive signals. Faces with neutral expressions signal a variety of socially relevant information, such that inferences about the big five personality traits, including Neuroticism, Extraversion and Agreeableness, can be accurately made on the basis of emotionally neutral facial photographs. Given the close link between Neuroticism and psychological distress, we investigated the effects of diminished central serotonin activity (achieved by tryptophan depletion) upon the accuracy of 52 healthy (non-clinical) adults' discriminations of personality from facial characteristics. All participants were able to discriminate reliably four of the big five traits. However, the tryptophan-depleted participants were specifically less accurate in discriminating Neuroticism than the matched non-depleted participants. These data suggest that central serotonin activity modulates the identification of not only negative facial emotional expression but also a broader class of signals about personality characteristics linked to psychological distress.


The Stanford Health Assessment Questionnaire: a review of its history, issues, progress, and documentation.
Over the last 2 decades, assessment of patient health status has undergone a dramatic paradigm shift, evolving from a predominant reliance on biochemical and physical measurements to an emphasis upon health outcomes based on the patient's personal appreciation of their illness. The Health Assessment Questionnaire (HAQ), published in 1980, was among the first instruments based on patient centered dimensions. The HAQ was designed to represent a model of patient oriented outcome assessment and has played a major role in diverse areas such as prediction of successful aging, inversion of the therapeutic pyramid in rheumatoid arthritis (RA), quantification of nonsteroidal antiinflammatory drug gastropathy, development of risk factor models for osteoarthrosis, and examination of mortality risks in RA. The HAQ has established itself as a valuable, effective, and sensitive tool for measurement of health status. It has increased the credibility and use of validated self-report measurement techniques as a quantifiable set of hard data endpoints and has contributed to a new appreciation of outcome assessment. We review the development, content, and dissemination of the HAQ and provide reference sources for its uses, translations, and validations. We discuss contemporary issues regarding outcome assessment instruments relative to the HAQ's identity and utility. These include: (1) the issue of labeling instruments as generic versus disease-specific; (2) floor and ceiling effects in scales such as "disability"; (3) distances between values on scales; and (4) the continuing introduction of new measurement instruments and their potential effects.


Seven challenges for modelling indirect transmission: vector-borne diseases, macroparasites and neglected tropical diseases.
Many of the challenges which face modellers of directly transmitted pathogens also arise when modelling the epidemiology of pathogens with indirect transmission--whether through environmental stages, vectors, intermediate hosts or multiple hosts. In particular, understanding the roles of different hosts, how to measure contact and infection patterns, heterogeneities in contact rates, and the dynamics close to elimination are all relevant challenges, regardless of the mode of transmission. However, there remain a number of challenges that are specific and unique to modelling vector-borne diseases and macroparasites. Moreover, many of the neglected tropical diseases which are currently targeted for control and elimination are vector-borne, macroparasitic, or both, and so this article includes challenges which will assist in accelerating the control of these high-burden diseases. Here, we discuss the challenges of indirect measures of infection in humans, whether through vectors or transmission life stages and in estimating the contribution of different host groups to transmission. We also discuss the issues of "evolution-proof" interventions against vector-borne disease.


Manifestations of personality in Online Social Networks: self-reported Facebook-related behaviors and observable profile information.
Despite the enormous popularity of Online Social Networking sites (OSNs; e.g., Facebook and Myspace), little research in psychology has been done on them. Two studies examining how personality is reflected in OSNs revealed several connections between the Big Five personality traits and self-reported Facebook-related behaviors and observable profile information. For example, extraversion predicted not only frequency of Facebook usage (Study 1), but also engagement in the site, with extraverts (vs. introverts) showing traces of higher levels of Facebook activity (Study 2). As in offline contexts, extraverts seek out virtual social engagement, which leaves behind a behavioral residue in the form of friends lists and picture postings. Results suggest that, rather than escaping from or compensating for their offline personality, OSN users appear to extend their offline personalities into the domains of OSNs.


Fragments of bacterial endoglycosidase s and immunoglobulin g reveal subdomains of each that contribute to deglycosylation.
Endoglycosidase S (EndoS) is a glycoside-hydrolase secreted by the bacterium Streptococcus pyogenes. EndoS preferentially hydrolyzes the N-linked glycans from the Fc region of IgG during infection. This hydrolysis impedes Fc functionality and contributes to the immune evasion strategy of S. pyogenes. Here, we investigate the mechanism of human serum IgG deactivation by EndoS. We expressed fragments of IgG1 and demonstrated that EndoS was catalytically active against all of them including the isolated CH2 domain of the Fc domain. Similarly, we sought to investigate which domains within EndoS could contribute to activity. Bioinformatics analysis of the domain organization of EndoS confirmed the previous predictions of a chitinase domain and leucine-rich repeat but also revealed a putative carbohydrate binding module (CBM) followed by a C-terminal region. Using expressed fragments of EndoS, circular dichroism of the isolated CBM, and a CBM-C-terminal region fusion revealed folded domains dominated by β sheet and α helical structure, respectively. Nuclear magnetic resonance analysis of the CBM with monosaccharides was suggestive of carbohydrate binding functionality. Functional analysis of truncations of EndoS revealed that, whereas the C-terminal of EndoS is dispensable for activity, its deletion impedes the hydrolysis of IgG glycans.


Effect of biofilm on the repair bond strengths of composites.
Composite restorations degrade during wear, but it is unknown how wear affects the composite surface and influences composite-to-composite bonding in minimally invasive repair. Here, it is hypothesized that in vitro exposure of composites to oral biofilm yields clinically relevant degradation of composite surfaces, and its influence on composite-to-composite bonding is determined. Biofilms on composite surfaces in vitro increased their roughness and decreased filler particle exposure, except for a microhybrid composite, similar to effects of clinical wear in palatal appliances. Failure shear stresses after intermediate-adhesive-resin application were significantly lower after aging by in vitro exposure to biofilms, while silica-coating maintained the same failure stress levels as in non-aged composites. Failure modes were predominantly cohesive after silica-coating, while intermediate-adhesive-resin application yielded more adhesive failure. It is concluded that in vitro exposure to oral biofilm is a clinically relevant aging condition, and that silica-coating is to be preferred for the repair of aged composites.


Stimulation of melanogenesis by tetradecanoylphorbol 13-acetate (TPA) in mouse melanocytes and neural crest cells.
In vitro studies have shown that the phorbol ester, 12-tetradecanoylphorbol 13-acetate (TPA) induces neural crest cell differentiation into melanocytes, and stimulates proliferation and differentiation of normal melanocytes. As TPA is not a physiological agent, its action is clearly mimicking some in vivo pathway involved in these processes. An understanding of the effect of TPA on the expression of melanogenic genes will therefore provide valuable insight into the molecular mechanisms regulating melanocyte differentiation. In this study, we utilized primary cultures of neural crest cells and an immortalized melanocyte cell line (DMEL-2) which proliferates in the absence of TPA, to explore the effects of TPA on key melanogenic effectors. In neural crest cells, TPA was found to be necessary for both microphthalmia associated transcription factor (Mitf) up-regulation and for melanin synthesis. Using northern blots, we show that in DMEL-2 cells, TPA significantly increases the messenger ribonucleic acid (mRNA) levels of the tyrosinase gene family (tyrosinase, Tyrp1 and Dct) and the expression of Mitf. Western blots demonstrate that in these TPA-treated cells there is a concomitant increase in Tyr, Tyrp1 and glycosylated Dct protein levels. Pax3, a known Mitf regulator, is unaltered by TPA treatment. This study demonstrates the utility of a novel cell line for investigating the long-term effects of TPA on melanogenesis and provides an understanding of how TPA enhances mouse melanocyte differentiation.


"Sometimes we can't fix things": a qualitative study of health care professionals' perceptions of end of life care for patients with heart failure.
Although heart failure has a worse prognosis than some cancers, patients often have restricted access to well-developed end of life (EoL) models of care. Studies show that patients with advanced heart failure may have a poor understanding of their condition and its outcome and, therefore, miss opportunities to discuss their wishes for EoL care and preferred place of death. We aimed to explore the perceptions and experiences of health care professionals (HCPs) working with patients with heart failure around EoL care. A qualitative in-depth interview study nested in a wider ethnographic study of unplanned admissions in patients with heart failure (HoldFAST). We interviewed 24 HCPs across primary, secondary and community care in three locations in England, UK - the Midlands, South Central and South West. The study revealed three issues impacting on EoL care for heart failure patients. Firstly, HCPs discussed approaches to communicating with patients about death and highlighted the challenges involved. HCPs would like to have conversations with patients and families about death and dying but are aware that patient preferences are not easy to predict. Secondly, professionals acknowledged difficulties recognising when patients have reached the end of their life. Lack of communication between patients and professionals can result in situations where inappropriate treatment takes place at the end of patients' lives. Thirdly, HCPs discussed the struggle to find alternatives to hospital admission for patients at the end of their life. Patients may be hospitalised because of a lack of planning which would enable them to die at home, if they so wished. The HCPs regarded opportunities for patients with heart failure to have ongoing discussions about their EoL care with clinicians they know as essential. These key professionals can help co-ordinate care and support in the terminal phase of the condition. Links between heart failure teams and specialist palliative care services appear to benefit patients, and further sharing of expertise between teams is recommended. Further research is needed to develop prognostic models to indicate when a transition to palliation is required and to evaluate specialist palliative care services where heart failure patients are included.


Prevalence of obesity was related to HLA-DQ in 2-4-year-old children at genetic risk for type 1 diabetes.
Body size is postulated to modulate type 1 diabetes as either a trigger of islet autoimmunity or an accelerator to clinical onset after seroconversion. As overweight and obesity continue to rise among children, the aim of this study was to determine whether human leukocyte antigen DQ (HLA-DQ) genotypes may be related to body size among children genetically at risk for type 1 diabetes. Repeated measures of weight and height were collected from 5969 children 2-4 years of age enrolled in The Environmental Determinants of Diabetes in the Young prospective study. Overweight and obesity was determined by the International Obesity Task Force cutoff values that correspond to body mass index (BMI) of 25 and 30 kg m(-)(2) at age 18. The average BMI was comparable across specific HLA genotypes at every age point. The proportion of overweight was not different by HL A, but percent obesity varied by age with a decreasing trend among DQ2/8 carriers (P for trend=0.0315). A multivariable regression model suggested DQ2/2 was associated with higher obesity risk at age 4 (odds ratio, 2.41; 95% confidence interval, 1.21-4.80) after adjusting for the development of islet autoantibody and/or type 1 diabetes. The HLA-DQ2/2 genotype may predispose to obesity among 2-4-year-old children with genetic risk for type 1 diabetes.


Performance of surface-plasma-wave fiber-optic polarizers.
Variations in the TM-like polarization transmission characteristics and polarization extinction ratio of surface-plasma-wave fiber-optic polarizers owing to variations in the depth of polishing and the metal film thickness are presented. The two effects have been isolated in carefully designed experiments and studied separately.


Dual-gene, dual-cell type therapy against an excitotoxic insult by bolstering neuroenergetics.
Increasing evidence suggests that glutamate activates the generation of lactate from glucose in astrocytes; this lactate is shuttled to neurons that use it as a preferential energy source. We explore this multicellular "lactate shuttle" with a novel dual-cell, dual-gene therapy approach and determine the neuroprotective potential of enhancing this shuttle. Viral vector-driven overexpression of a glucose transporter in glia enhanced glucose uptake, lactate efflux, and the glial capacity to protect neurons from excitotoxicity. In parallel, overexpression of a lactate transporter in neurons enhanced lactate uptake and neuronal resistance to excitotoxicity. Finally, overexpression of both transgenes in the respective cell types provided more protection than either therapy alone, demonstrating that a dual-cell, dual-gene therapy approach gives greater neuroprotection than the conventional single-cell, single-gene strategy.


Luc's abscess--a rare complication of middle-ear infection.
A case of subperiosteal temporal abscess of otitic origin is presented. This is an unusual complication of otitis media. The pathogenesis of Luc's abscess is different from other extracranial complications of middle-ear infections in that it is not associated with mastoid infection which results in subperiosteal pus formation. Based on our experience and the reports from the turn of the century, we present the presumptive pathogenesis and clinical features. We contend that these patients run an unexpectedly benign course, and require concomitantly more conservative treatment than other otitic abscesses.